CN1827764A - Degradation bacteria for carbendazim pesticide residue and bacterial agent produced thereby - Google Patents
Degradation bacteria for carbendazim pesticide residue and bacterial agent produced thereby Download PDFInfo
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- CN1827764A CN1827764A CNA2005100225468A CN200510022546A CN1827764A CN 1827764 A CN1827764 A CN 1827764A CN A2005100225468 A CNA2005100225468 A CN A2005100225468A CN 200510022546 A CN200510022546 A CN 200510022546A CN 1827764 A CN1827764 A CN 1827764A
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Abstract
The invention provides a carbendazim pesticide residue degrading bacteria and a prodegradant, which belongs to biological high-tech domain. The used bacterial strain DJL-6 is judged as Rhodococcus sp. The main biological feature is: G+, the thallus is of ball rod and atrichosis; the reaction between catalase and V-P is positive; the oxidase reaction is negative; the barley sugar, carubinose, citrate, acetamide and agedoite can be used as unique carbon source or nitrogen source to carry out its growth, but the lactin and bitter almond acid can not be used as unique carbon source to carry out its growth; and the bacterial colony is orange when cultivated on LB culture medium. The pesticide residual quantity of the crop can be reduced by more than 95 % by directly applying the degrading bacteria product, which solves the overproof pesticide residue problem, and can produce non-toxic and nuisanceless environmental farm product.
Description
One, technical field
The microbial inoculum of a kind of derosal pesticide residual degrading bacteria of the present invention and production thereof belongs to biological high-tech field, is to utilize method of microorganism degraded chemical pesticide residual, is applicable to the production and the processing of green non-polluted farm product in the modern agriculture production.
Two, technical background
The use of chemical pesticide has promoted modern agricultural development widely, for the loss that reasons such as a large amount of due to illness insect pests cause has been retrieved in agriculture production, and greatly improved the efficiency and the mechanization degree of agriculture production, in the modernization of agriculture production, played immeasurable effect.But agricultural chemicals all is some toxic and harmful substances simultaneously, after they enter ecotope, has not only killed the deleterious biology to the mankind, also the ecosystem has been caused serious harm simultaneously.A large amount of uses of agricultural chemicals cause that pesticide residue exceed standard in the agricultural-food, have had a strong impact on the foreign exchange earning of people health and agricultural-food.Along with the raising of people's living standard and the enhancing of environmental protection consciousness, the whole society also more and more pays close attention to direct harm and potential impact that pesticide residue are brought, and people press for the green agricultural product of the cleaning of non agricultural chemical residuum pollution.But agricultural form and demographic situation from present China, use to stop agricultural chemicals that to reduce farm output be impracticable as the nuisanceless production model of cost, agricultural chemicals also will continue to bring into play enormous function as the effective means of control disease and pest in agriculture production.How solving this double-barreled question in the agriculture production just becomes the research focus of each subject.Laboratory and real application research all show pesticide residue " storehouse " capacity that utilizes microorganism to degrade to eliminate in the soil, thereby reduce the pesticide residue in the agricultural-food such as grain, vegetables, tealeaves, and the quality and the economic worth that improve agricultural-food are feasible.
Three, summary of the invention
Technical problem the objective of the invention is at practical problems in the production practice and demand, develops out a kind of novel bacterium for degradating residual agricultural chemical, uses this microbial inoculum can make the residual quantity of derosal reduce more than 90%.
Be main contents of the present invention below the technical scheme:
The invention provides a kind of degradation bacteria of eliminating the derosal pesticide residue, its bacterial strain is that a strain gram positive bacterium DJL-6 (was deposited at Chinese microorganism strain and preserves management committee common micro-organisms center on October 12nd, 2005, culture presevation number is CGMCC NO.1478), through being accredited as Rhod (Rhodococcus sp.).Main biological characteristics is: G
+, thalline is the club shape, does not have flagellum; Catalase and V-P reaction are positive; The oxydase reaction feminine gender; Can utilize maltose, seminose, Citrate trianion, ethanamide and l-asparagine to grow for sole carbon or nitrogenous source, can not be that sole carbon source is grown with lactose, mandelic acid; Bacterium colony is orange when cultivating on the LB substratum, can not utilize Sodium Benzoate to grow for sole carbon source.The Genbank number of landing of its 16S rDNA is DQ090961.
The technology of using above-mentioned derosal pesticide residual degrading bacteria to produce microbial inoculum is: inclined-plane kind-shake bottle a kinds-seeding tank-production jar-product (pack formulation and be liquid bacterial agent or solid absorption microbial inoculum).
Detailed implementation step of the present invention is:
1) the test tube kind with derosal pesticide residual degrading bacteria DJI-6 is inoculated in the LB substratum, and shaking culture is to logarithmic phase;
2) above-mentioned cultured bacterial classification is inoculated into 500 liters of seeding tanks by 10% inoculum size of seed tank culture base, be cultured to logarithmic phase, the used culture medium prescription of seeding tank is: glucose 0.8%, (NH
4)
2SO
41%, K
2HPO
40.2%, MgSO
40.05%, NaCl 0.01%, CaCO
30.3%, yeast extract paste 0.02%, pH value 7.2-7.5;
3) seed liquor is produced a jar cultivation by the 10% inoculum size access of production jar substratum, it is identical with the seed tank culture base to produce the used substratum of jar;
The air flow of sterile air is 1 in the culturing process of seeding tank and production jar: 0.6-1.2, stirring velocity is 180-240 rev/min, culture temperature is 30-35 ℃, the whole process incubation time is 48-60 hour, after the fermentation ends thalline quantity reach 1,000,000,000/more than the ml, fermentation is finished the back nutrient solution and is gone out jar and directly be distributed into liquid dosage form with plastic barrel or packing bottle or adopt peat to adsorb and be distributed into the solid fungicide formulation with packing bag.
Beneficial effect
It is low, easy to use that the bacterium for degradating residual agricultural chemical that uses this invention to produce has the production use cost, and the advantage that removal effect is good is adapted at national grain oil ﹠ vegetable production export base or has the local big area of green food brand mark to promote the use of.
The present invention is for preserving the ecological environment, and the protection people health improves additional value of farm products and has great importance.Using this degradation bacterial agent normally to use chemical pesticide to carry out the prevention and control of plant diseases, pest control in the normal growth process of farm crop and guaranteeing that the agricultural chemicals residual content meets green food in the agricultural-food requires degradation bacteria DJL-6 to have higher degradation efficiency, the residual quantity degraded that can make the derosal agricultural chemicals is more than 90%, greatly reduce the workload in production and the use, reduced production and use cost.
Use this degradation bacterial agent in the normal growth process of farm crop, normally to use chemical pesticide to carry out the prevention and control of plant diseases, pest control and guarantee that the agricultural chemicals residual content meets the green food requirement in the agricultural-food.And this bacterium is a gram-positive microorganism, the effect that agricultural chemicals uses after the survival time in field suitably can not influence.
The present invention has successfully solved in the agriculture production derosal pesticide residue problem that exceeds standard, and has both given full play to the efficiently fast effect of chemical pesticide in plant pest management, can produce non-toxic and non-pollution green agricultural product again.
Four, embodiment
Below narrate embodiments of the invention.
The original seed of degradation bacteria DJL-6 of the present invention is activated on culture dish, and measure degradation property, be inoculated on the test tube slant standby.The test tube kind is inoculated in 1000ml and shakes in the bottle, and the LB substratum of 200ml is added in the inside, and constant-temperature shaking culture is prepared the inoculation seeding tank to logarithmic phase.500 liters of seeding tanks, 400 liters of charging capacitys, culture medium prescription is: glucose 0.8%, (NH
4)
2SO
41%, K
2HPO
40.2%, MgSO
40.05%, NaCl 0.01%, CaCO
30.3%, yeast extract paste 0.02%, pH value 7.2-7.5.The back 121 ℃ of high pressure moist heat sterilizations that finish that feed intake, be cooled to 33 ℃ after, the above-mentioned cultured bottle bacterial classification that shakes is inoculated into 500 liters of seeding tanks by 10% inoculum size, be cultured to logarithmic phase, stirring velocity is 220 rev/mins, sterile air feeding amount is 1: 0.8.The seed liquor that arrives logarithmic phase is produced jar by 10% inoculum size access cultivate, it is identical with the seed tank culture base to produce the used medium component of jar.Produce 5 tons of tankages, 4.5 tons of charging capacitys.Production jar 1.1kg/cm after feeding intake
2Pressure under, 121 ℃ of high pressure moist heat sterilizations, below the sterilization postcooling to 35 ℃, logical sterile air keeps sterile state standby.A postvaccinal production jar temperature is controlled at 30-35 ℃, and the air flow of sterile air is 1 in the culturing process of production jar: 0.8-1.0, and stirring velocity is 240 rev/mins, the whole process flow incubation time is 48-60 hour.After the fermentation ends thalline quantity reach 1,000,000,000/more than the ml.
Fermentation is finished the back nutrient solution and is gone out jar and directly be distributed into liquid dosage form with plastic barrel or packing bottle or adopt peat to adsorb and be distributed into the solid fungicide formulation with packing bag.
Product with the aforesaid method gained carries out chemical residual degradation experiment and demonstrations such as soil, water body.After soil or water body were used the derosal agricultural chemicals, above-mentioned microbial inoculum kept 3 days by the using method requirement in soil or water body after, get an amount of soil or water body after extracting, with the residual content of derosal in liquid chromatography for measuring soil or the vegetables (measuring), from the result, find out the remains of pesticide (table 1) that after degradation bacteria is handled, can reduce greatly in soil and the water body by Shanxi Province's agricultural chemicals key lab (Taiyuan).
Derosal residue detection result in table 1 bacterial strain DJL-6 degraded soil and the water body
| The sample title | The former numbering of sample | Derosal content (mgkg -1) | Detection limit | Clearance (%) | |
| Water body | DJL-6 | Contrast | 5.57 | 1.0 | 100％ |
| Handle | Do not detect | 1.0 | 100％ | ||
| Soil | DJL-6 | Contrast | 20.48 | 0.5 | 100％ |
| Handle | Do not detect | 0.5 | 100％ | ||
Examining report
Examining report numbering: 0415
| Client | Life science institute of Agricultural University Of Nanjing department of microbiology | Consignor and address | Xu Jingliang | ||||
| The unit of being examined | —— | Contact person and address | —— | ||||
| The sample date | 2004.9.21, September 24 dark freezing | The consignment agreement numbering | 0415 | ||||
| Come sample loading mode | Mailing | Detect the date | 11.11-11.22 | ||||
| Sample state | Water sample, soil sample | Detection method | The retention analysis of derosal in soil and substratum | ||||
| Sample detection result | |||||||
| The sample title | The former numbering of sample | Test item | Detected result (mg/kg) | Detection limit (mg/kg) | |||
| Water | DJL-6 handles | Derosal | Do not detect | 1.0 | |||
| Soil | DJL-6 handles 1 | Derosal | Do not detect | 0.5 | |||
| Soil | DJL-6 handles 2 | Derosal | Do not detect | 0.5 | |||
| Soil | DJL-6 handles 3 | Derosal | Do not detect | 0.5 | |||
| Water | DJL-6-2 handles | Derosal | Do not detect | 1.0 | |||
| Soil | DJL-6-2 handles 1 | Derosal | Do not detect | 0.5 | |||
| Soil | DJL-6-2 handles 2 | Derosal | Do not detect | 0.5 | |||
| Soil | DJL-6-2 handles 3 | Derosal | Do not detect | 0.5 | |||
| Water | CK | Derosal | 5.57 | 1.0 | |||
| Soil | CK contrast 1 | Derosal | 20.48 | 0.5 | |||
| Soil | CK contrast 2 | Derosal | 19.15 | 0.5 | |||
| Remarks: | |||||||
Establishment: Qin Shu audit: Wang Xia signed by (post, signature): Laboratory Chief Qiao Xiong Chinese parasol tree
The date of issuance: on November 26th, 2004
Shanxi Province agricultural chemicals key lab
Claims (2)
1. derosal pesticide residual degrading bacteria, it is characterized in that, this microorganism is the bacterial strain DJL-6 of the Rhod (Rhodococcus sp.) of Gram-positive, be deposited at Chinese microorganism strain on October 12nd, 2005 and preserve management committee common micro-organisms center, culture presevation number is CGMCC NO.1478, and main biological characteristics is: G
+, thalline is the club shape, does not have flagellum; Catalase and V-P reaction are positive; The oxydase reaction feminine gender; Can utilize maltose, seminose, Citrate trianion, ethanamide and l-asparagine to grow for sole carbon or nitrogenous source, can not be that sole carbon source is grown with lactose, mandelic acid; Bacterium colony is orange when cultivating on the LB substratum, can not utilize Sodium Benzoate to grow for sole carbon source, and the Genbank number of landing of its 16S rDNA is DQ090961.
2. bacterium for degradating residual agricultural chemical of producing with the described derosal pesticide residual degrading bacteria of claim 1 is to produce by the following method:
1) the test tube kind with derosal pesticide residual degrading bacteria DJL-6 is inoculated in the LB substratum, and shaking culture is to logarithmic phase;
2) above-mentioned cultured bacterial classification is inoculated into 500 liters of seeding tanks by 10% inoculum size of seed tank culture base, be cultured to logarithmic phase, the used culture medium prescription of seeding tank is: glucose 0.8%, (NH
4)
2SO
41%, K
2HPO
40.2%, MgSO
40.05%, NaCl 0.01%, CaCO
30.3%, yeast extract paste 0.02%, pH value 7.2-7.5;
3) seed liquor is produced a jar cultivation by the 10% inoculum size access of production jar substratum, it is identical with the seed tank culture base to produce the used substratum of jar;
4) air flow of sterile air is 1 in the culturing process of seeding tank and production jar: 0.6-1.2, stirring velocity is 180-240 rev/min, culture temperature is 30-35 ℃, the whole process incubation time is 48-60 hour, after the fermentation ends thalline quantity reach 1,000,000,000/more than the ml, fermentation is finished the back nutrient solution and is gone out jar and directly be distributed into liquid dosage form with plastic barrel or packing bottle or adopt peat to adsorb and be distributed into the solid fungicide formulation with packing bag.
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| CNB2005100225468A CN100339473C (en) | 2005-12-23 | 2005-12-23 | Degradation bacteria for carbendazim pesticide residue and bacterial agent produced thereby |
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| CN100339473C CN100339473C (en) | 2007-09-26 |
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Cited By (6)
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|---|---|---|---|---|
| CN101649306B (en) * | 2009-09-15 | 2012-05-23 | 山东大学 | Enzyme capable of degrading pesticide carbendazim and encoding gene and application thereof |
| CN104862245A (en) * | 2015-04-16 | 2015-08-26 | 西北农林科技大学 | Carbendazim degrading bacteria MBC-6F and application thereof |
| CN105820982A (en) * | 2016-04-29 | 2016-08-03 | 河北省科学院生物研究所 | Rhodococcus ruber XS-2 strain and biological microbial inoculant, and preparation method and application thereof |
| CN108070605A (en) * | 2018-01-05 | 2018-05-25 | 南京农业大学 | Carbendazim degrading enzyme CbmA and its encoding gene and application |
| CN110484449A (en) * | 2019-07-30 | 2019-11-22 | 南京农业大学 | A kind of protective agent of carbendazim degrading bacterium and its preparation method and application |
| CN110747150A (en) * | 2019-12-05 | 2020-02-04 | 中国农业科学院烟草研究所 | Carbendazim degrading bacterium and cultivation method and application thereof |
Family Cites Families (2)
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| CN1242053C (en) * | 2004-01-08 | 2006-02-15 | 南京农业大学 | Carbofuran pesticide residual degraded bacteria and its produced bacterial agent |
| CN1563356A (en) * | 2004-03-24 | 2005-01-12 | 南京农业大学 | Bacterium of degrading residual of organophorus pespared agent of bacterium |
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2005
- 2005-12-23 CN CNB2005100225468A patent/CN100339473C/en not_active Expired - Fee Related
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101649306B (en) * | 2009-09-15 | 2012-05-23 | 山东大学 | Enzyme capable of degrading pesticide carbendazim and encoding gene and application thereof |
| CN104862245A (en) * | 2015-04-16 | 2015-08-26 | 西北农林科技大学 | Carbendazim degrading bacteria MBC-6F and application thereof |
| CN104862245B (en) * | 2015-04-16 | 2017-09-19 | 西北农林科技大学 | One plant of carbendazim degrading bacterium MBC 6F and its application |
| CN105820982A (en) * | 2016-04-29 | 2016-08-03 | 河北省科学院生物研究所 | Rhodococcus ruber XS-2 strain and biological microbial inoculant, and preparation method and application thereof |
| CN105820982B (en) * | 2016-04-29 | 2019-06-28 | 河北省科学院生物研究所 | A kind of Rhodococcus ruber XS-2, its bacteria agent and preparation method and application |
| CN108070605A (en) * | 2018-01-05 | 2018-05-25 | 南京农业大学 | Carbendazim degrading enzyme CbmA and its encoding gene and application |
| CN108070605B (en) * | 2018-01-05 | 2021-06-01 | 南京农业大学 | Carbendazim degrading enzyme CbmA as well as coding gene and application thereof |
| CN110484449A (en) * | 2019-07-30 | 2019-11-22 | 南京农业大学 | A kind of protective agent of carbendazim degrading bacterium and its preparation method and application |
| CN110747150A (en) * | 2019-12-05 | 2020-02-04 | 中国农业科学院烟草研究所 | Carbendazim degrading bacterium and cultivation method and application thereof |
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| CN100339473C (en) | 2007-09-26 |
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Assignee: Yixing Qiancheng Biological Co., Ltd. Assignor: Nanjing Agricultural University Contract record no.: 2010320000457 Denomination of invention: Degradation bacteria for carbendazim pesticide residue and bacterial agent produced thereby Granted publication date: 20070926 License type: Exclusive License Open date: 20060906 Record date: 20100426 |
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